Aim: The aim of this research is to determine the effect of acute hyperbaric oxygenation on oxidative stress parameters in healthy Sprague-Dawley rats. Materials and methods: Healthy male rats aged 9-11 weeks of age were divided into two groups: CTRL, control group (healthy untreated rats) and A-HBO2 group (rats exposed to hyperbaric oxygen in hyperbaric chamber (100% oxygen, 2 bar / 2 hours ). The rats were anesthetized by a combination of ketamine (75 mg / kg) and midazolam (2.5 mg / kg). Blood samples were taken, in which the oxidative stress indicators were measured, using TBARS and FRAP methods. Also, body mass and blood pressure were measured in each animal. Results: There weren't significant differences in the body weight of the rat, while blood pressure was significantly reduced in the A-HBO2 group compared to the control group. TBARS was significantly increased in the A-HBO2 group compared to the control group, while the antioxidant capacity among the investigated groups did not significantly change. Conclusion: Acute exposure to hyperbaric oxygenation increases oxidative stress in healthy Spargue-Dawley rats.